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Research article
Zhenghong Li
Research Institute of Resources Insects, Chinese Academy of Forestry, 650224 Kunming, China
Corresponding Author
ORCiD: https://orcid.org/0000-0003-1677-4628
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This work is licensed under a CC BY 4.0 License. Read Full License
Background: Photoperiod-regulated floral transition is vital to the flowering plant. Luculia gratissima ‘Xiangfei’ is a flowering ornamental plant with high development potential and is a short-day woody perennial. However, the genetic regulation of short-day-induced floral transition in L. gratissima is unclear. To systematically research the responses of L. gratissima during this process, dynamic changes in morphology, physiology, and transcript levels were observed and identified in different developmental stages of long-day and short-day-treated shoot apexes.
Results: The results showed that floral transition in L. gratissima occurred 10 d after short-day induction, but flower bud differentiation did not occur under long-day conditions. A total of 1,226 differentially expressed genes were identified, of which 146 genes were associated with flowering pathways of sugar, phytohormones, photoperiod, ambient temperature, and aging signals, as well as floral integrator and meristem identity genes. The trehalose-6-phosphate signal positively modulated floral transition by interacting with SPL4 in the aging pathway. Endogenous gibberellin, abscisic acid, cytokinin, and jasmonic acid promoted floral transition, whereas strigolactone inhibited it. In the photoperiod pathway, FD, COL12, and NF-Ys positively controlled floral transition, whereas PRR7, FKF1, and LUX negatively regulated it. SPL4 and pEARLI1 positively affected floral transition. SOC1 and AGL24 integrated multiple flowering signals to modulate the expression of FUL/AGL8, AP1, LFY, SEPs, SVP, and TFL1, thereby regulating floral transition. Finally, we propose a regulatory network model for short-day-induced floral transition in L. gratissima.
Conclusions: Short-day photoperiod activated systemic responses of morphology, physiology, and transcript levels in L. gratissima and induced the generation of floral transition signals in the photoperiod pathway. Furthermore, multiple flowering signal pathways including phytohormone-, sugar-, temperature-, age-related genes synergistically control this process. This study improves our understanding of flowering time regulation in L. gratissima and provides knowledge for its production and commercialization.
Keywords
Luculia gratissima, Floral transition, Photoperiod, Flowering pathway, Phytohormone, Regulatory network
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This is a list of supplementary files associated with this preprint. Click to download.
- Additionalfile17TableS9.docx
Additional file 17: Table S9. List of primer sets used in the study.
- Additionalfile13TableS7.xlsx
Additional file 13: Table S7. Candidate floral differentially expressed genes (DEGs) uniquely or commonly belonging to each comparison. a Candidate floral DEGs uniquely belonging to each comparison; b Candidate floral DEGs commonly belonging to each comparison.
- Additionalfile12TableS6.xlsx
Additional file 12: Table S6. Gene ontology (GO) enrichment analysis of genes specific to each co-expression module identified by weighted gene co-expression network analysis (WGCNA) of Luculia gratissima. a Significantly enriched GO terms of genes specific to each co-expression module; b Element set of overlapping regions of significantly enriched GO terms in 11 co-expression modules.
- Additionalfile15Fig.S7.docx
Additional file 15: Fig. S7. Co-expression network of differentially expressed genes (DEGs) associated with floral transition in Luculia gratissima. A total of 126 floral transition-related DEGs with edge weight > 0.1 were mapped by Cytoscape. The larger and redder the circles, the higher is the connectivity. When more than two unigenes are the same gene, the unigene ID is marked in parentheses after the gene name.
- Additionalfile14TableS8.xlsx
Additional file 14: Table S8. Significance of candidate floral differentially expressed genes involved in various flowering pathways.
- Additionalfile11TableS5.xlsx
Additional file 11: Table S5. Co-expression module analysis for 1,226 differentially expressed genes at four developmental stages of Luculia gratissima. a The kME (module membership-measuring how correlated a gene is to the eigengene), kME-p-value, and connectivity (connectivity with all the other genes in the same module) of each gene in the 11 co-expression modules; b The correlation co-efficient between the module eigengene and the specific stage and p-value of the corresponding correlation co-efficient.
- Additionalfile10Fig.S6.docx
Additional file 10: Fig. S6. Cellular response overview in each comparison. Cellular response overview maps were designed by MapMan based on the transcript levels of differentially expressed genes (DEGs) in (a) LD7-vs.-SD7, (b) LD10-vs.-SD10, (c) LD13-vs.-SD13, and (d) LD19-vs.-SD19. The color indicates log2 value of fold changes, with green and red colors representing down- and up-regulated transcripts, respectively.
- Additionalfile9Fig.S5.docx
Additional file 9: Fig. S5. Metabolism overview in each comparison. The metabolism overview map was designed by MapMan based on the transcript levels of differentially expressed genes (DEGs) in (a) LD7-vs.-SD7, (b) LD10-vs.-SD10, (c) LD13-vs.-SD13, and (d) LD19-vs.-SD19. The color indicates log2 value of fold changes, with green and red colors representing down- and up-regulated transcripts, respectively.
- Additionalfile8Fig.S4.docx
Additional file 8: Fig. S4. Regulation overview in each comparison. The regulation overview map was designed by MapMan based on the transcript levels of differentially expressed genes (DEGs) within (a) LD7-vs.-SD7, (b) LD10-vs.-SD10, (c) LD13-vs.-SD13, and (d) LD19-vs.-SD19. The color indicates log2 value of fold changes, with green and red colors representing down- and up-regulated transcripts, respectively.
- Additionalfile7TableS4.xlsx
Additional file 7: Table S4. MapMan annotation of differentially expressed genes (DEGs) at four developmental stages of Luculia gratissima. a MapMan annotation of DEGs for comparison LD7-vs.-SD7; b MapMan annotation of DEGs for comparison LD10-vs.-SD10; c MapMan annotation of DEGs for comparison LD13-vs.-SD13; d MapMan annotation of DEGs for comparison LD19-vs.-SD19.
- Additionalfile16SupplementaryData.docx
Additional file 16: Supplementary Data. Evaluation of the optimal reference genes (Unpublished data).
- Additionalfile6TableS3.xlsx
Additional file 6: Table S3. Differentially expressed genes (DEGs) at four developmental stages of Luculia gratissima. a DEGs in the comparison LD7-vs.-SD7; b DEGs in the comparison LD10-vs.-SD10; c DEGs in the comparison LD13-vs.-SD13; d DEGs in the comparison LD19-vs.-SD19.
- Additionalfile5Fig.S3.docx
Additional file 5: Fig. S3. Differentially expressed genes (DEGs) in each comparison. Numbers of DGEs in comparisons (a) LD7-vs.-SD7, (b) LD10-vs.-SD10, (c) LD13-vs.-SD13, and (d) LD19-vs.-SD19. e Venn diagram showing differentially and stage-specific gene profile per comparison.
- Additionalfile4Fig.S2.docx
Additional file 4: Fig. S2. Relative expression of flowering-related genes at four time points under short-day and long-day treatments. The relative gene expression as detected by RNA sequencing (RNA-seq) and confirmed by quantitative real-time PCR (qRT-PCR).
- Additionalfile3Fig.S1.docx
Additional file 3: Fig. S1. Unigenes length distribution and annotation statistics. a Length distribution of the assembled unigenes. b Venn diagram for the number of unigenes annotated by six different databases – NR, KOG, KEGG, Swiss-Prot, eggNOG and Pfam. c KEGG classification of unigenes. d Gene ontology classification of unigenes.
- Additionalfile2TableS2.docx
Additional file 2: Table S2. Summary of the assembled unigenes.
- Additionalfile1TableS1.docx
Additional file 1: Table S1. RNA sequencing (RNA-seq) statistics.
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Research article
Zhenghong Li
Research Institute of Resources Insects, Chinese Academy of Forestry, 650224 Kunming, China
Corresponding Author
ORCiD: https://orcid.org/0000-0003-1677-4628
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
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CURRENT STATUS: Posted
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Posted 19 Oct, 2020
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License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Photoperiod-regulated floral transition is vital to the flowering plant. Luculia gratissima ‘Xiangfei’ is a flowering ornamental plant with high development potential and is a short-day woody perennial. However, the genetic regulation of short-day-induced floral transition in L. gratissima is unclear. To systematically research the responses of L. gratissima during this process, dynamic changes in morphology, physiology, and transcript levels were observed and identified in different developmental stages of long-day and short-day-treated shoot apexes.
Results: The results showed that floral transition in L. gratissima occurred 10 d after short-day induction, but flower bud differentiation did not occur under long-day conditions. A total of 1,226 differentially expressed genes were identified, of which 146 genes were associated with flowering pathways of sugar, phytohormones, photoperiod, ambient temperature, and aging signals, as well as floral integrator and meristem identity genes. The trehalose-6-phosphate signal positively modulated floral transition by interacting with SPL4 in the aging pathway. Endogenous gibberellin, abscisic acid, cytokinin, and jasmonic acid promoted floral transition, whereas strigolactone inhibited it. In the photoperiod pathway, FD, COL12, and NF-Ys positively controlled floral transition, whereas PRR7, FKF1, and LUX negatively regulated it. SPL4 and pEARLI1 positively affected floral transition. SOC1 and AGL24 integrated multiple flowering signals to modulate the expression of FUL/AGL8, AP1, LFY, SEPs, SVP, and TFL1, thereby regulating floral transition. Finally, we propose a regulatory network model for short-day-induced floral transition in L. gratissima.
Conclusions: Short-day photoperiod activated systemic responses of morphology, physiology, and transcript levels in L. gratissima and induced the generation of floral transition signals in the photoperiod pathway. Furthermore, multiple flowering signal pathways including phytohormone-, sugar-, temperature-, age-related genes synergistically control this process. This study improves our understanding of flowering time regulation in L. gratissima and provides knowledge for its production and commercialization.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
This is a list of supplementary files associated with this preprint. Click to download.
- Additionalfile17TableS9.docx
Additional file 17: Table S9. List of primer sets used in the study.
- Additionalfile13TableS7.xlsx
Additional file 13: Table S7. Candidate floral differentially expressed genes (DEGs) uniquely or commonly belonging to each comparison. a Candidate floral DEGs uniquely belonging to each comparison; b Candidate floral DEGs commonly belonging to each comparison.
- Additionalfile12TableS6.xlsx
Additional file 12: Table S6. Gene ontology (GO) enrichment analysis of genes specific to each co-expression module identified by weighted gene co-expression network analysis (WGCNA) of Luculia gratissima. a Significantly enriched GO terms of genes specific to each co-expression module; b Element set of overlapping regions of significantly enriched GO terms in 11 co-expression modules.
- Additionalfile15Fig.S7.docx
Additional file 15: Fig. S7. Co-expression network of differentially expressed genes (DEGs) associated with floral transition in Luculia gratissima. A total of 126 floral transition-related DEGs with edge weight > 0.1 were mapped by Cytoscape. The larger and redder the circles, the higher is the connectivity. When more than two unigenes are the same gene, the unigene ID is marked in parentheses after the gene name.
- Additionalfile14TableS8.xlsx
Additional file 14: Table S8. Significance of candidate floral differentially expressed genes involved in various flowering pathways.
- Additionalfile11TableS5.xlsx
Additional file 11: Table S5. Co-expression module analysis for 1,226 differentially expressed genes at four developmental stages of Luculia gratissima. a The kME (module membership-measuring how correlated a gene is to the eigengene), kME-p-value, and connectivity (connectivity with all the other genes in the same module) of each gene in the 11 co-expression modules; b The correlation co-efficient between the module eigengene and the specific stage and p-value of the corresponding correlation co-efficient.
- Additionalfile10Fig.S6.docx
Additional file 10: Fig. S6. Cellular response overview in each comparison. Cellular response overview maps were designed by MapMan based on the transcript levels of differentially expressed genes (DEGs) in (a) LD7-vs.-SD7, (b) LD10-vs.-SD10, (c) LD13-vs.-SD13, and (d) LD19-vs.-SD19. The color indicates log2 value of fold changes, with green and red colors representing down- and up-regulated transcripts, respectively.
- Additionalfile9Fig.S5.docx
Additional file 9: Fig. S5. Metabolism overview in each comparison. The metabolism overview map was designed by MapMan based on the transcript levels of differentially expressed genes (DEGs) in (a) LD7-vs.-SD7, (b) LD10-vs.-SD10, (c) LD13-vs.-SD13, and (d) LD19-vs.-SD19. The color indicates log2 value of fold changes, with green and red colors representing down- and up-regulated transcripts, respectively.
- Additionalfile8Fig.S4.docx
Additional file 8: Fig. S4. Regulation overview in each comparison. The regulation overview map was designed by MapMan based on the transcript levels of differentially expressed genes (DEGs) within (a) LD7-vs.-SD7, (b) LD10-vs.-SD10, (c) LD13-vs.-SD13, and (d) LD19-vs.-SD19. The color indicates log2 value of fold changes, with green and red colors representing down- and up-regulated transcripts, respectively.
- Additionalfile7TableS4.xlsx
Additional file 7: Table S4. MapMan annotation of differentially expressed genes (DEGs) at four developmental stages of Luculia gratissima. a MapMan annotation of DEGs for comparison LD7-vs.-SD7; b MapMan annotation of DEGs for comparison LD10-vs.-SD10; c MapMan annotation of DEGs for comparison LD13-vs.-SD13; d MapMan annotation of DEGs for comparison LD19-vs.-SD19.
- Additionalfile16SupplementaryData.docx
Additional file 16: Supplementary Data. Evaluation of the optimal reference genes (Unpublished data).
- Additionalfile6TableS3.xlsx
Additional file 6: Table S3. Differentially expressed genes (DEGs) at four developmental stages of Luculia gratissima. a DEGs in the comparison LD7-vs.-SD7; b DEGs in the comparison LD10-vs.-SD10; c DEGs in the comparison LD13-vs.-SD13; d DEGs in the comparison LD19-vs.-SD19.
- Additionalfile5Fig.S3.docx
Additional file 5: Fig. S3. Differentially expressed genes (DEGs) in each comparison. Numbers of DGEs in comparisons (a) LD7-vs.-SD7, (b) LD10-vs.-SD10, (c) LD13-vs.-SD13, and (d) LD19-vs.-SD19. e Venn diagram showing differentially and stage-specific gene profile per comparison.
- Additionalfile4Fig.S2.docx
Additional file 4: Fig. S2. Relative expression of flowering-related genes at four time points under short-day and long-day treatments. The relative gene expression as detected by RNA sequencing (RNA-seq) and confirmed by quantitative real-time PCR (qRT-PCR).
- Additionalfile3Fig.S1.docx
Additional file 3: Fig. S1. Unigenes length distribution and annotation statistics. a Length distribution of the assembled unigenes. b Venn diagram for the number of unigenes annotated by six different databases – NR, KOG, KEGG, Swiss-Prot, eggNOG and Pfam. c KEGG classification of unigenes. d Gene ontology classification of unigenes.
- Additionalfile2TableS2.docx
Additional file 2: Table S2. Summary of the assembled unigenes.
- Additionalfile1TableS1.docx
Additional file 1: Table S1. RNA sequencing (RNA-seq) statistics.
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